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Related Experiment Videos

Analysis of multiple Invs transcripts in mouse and MDCK cells.

Heather H Ward1, Jinghong Wang, Carriel Phillips

  • 1Department of Pathology, Indiana University School of Medicine, 950 West Walnut, RII 202, Indianapolis, IN 46202, USA.

Genomics
|November 10, 2004
PubMed
Summary

Researchers identified multiple inversin (INVS) splice variants in mouse and canine kidneys. These variants, caused by exon skipping, alter inversin protein motifs crucial for kidney development and left-right asymmetry.

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Area of Science:

  • Developmental Biology
  • Genetics
  • Molecular Biology

Background:

  • Infantile nephronophthisis is linked to cystic kidneys, situs inversus, and INVS mutations.
  • The function of inversin (INVS), a protein product of the INVS gene, is not fully understood.
  • Evidence suggests multiple inversin isoforms exist with variations in molecular weight, cellular localization, and binding partners.

Purpose of the Study:

  • To identify alternative INVS transcripts and understand their role in kidney development.
  • To investigate how INVS splice variants affect inversin protein structure and function.
  • To determine the contribution of specific inversin protein motifs to left-right asymmetry and kidney development.

Main Methods:

  • Northern blot analysis of normal mouse kidney RNA.

Related Experiment Videos

  • Reverse transcription-polymerase chain reaction (RT-PCR) to detect Invs transcripts with exon skipping.
  • Sequence analysis of canine INVS and comparison with mouse and human orthologs.

    • Main Results:

    • Northern blots revealed four distinct bands for Invs cDNA in mouse kidney.
    • RT-PCR identified Invs transcripts in mouse kidney with skipping of exons 5, 11, or 13.
    • Canine INVS shares significant sequence identity with mouse (74%) and human (84%) inversin and produces a transcript skipping exon 12.
    • Exon skipping in INVS leads to the loss of critical protein motifs, including ankyrin repeats, IQ domains, destruction boxes, and nuclear localization signals.

    Conclusions:

    • The study identified novel INVS splice variants in both mouse and canine models.
    • These splice variants result in altered inversin protein isoforms, impacting key functional domains.
    • Understanding these INVS splice variants is crucial for elucidating their role in left-right body asymmetry and kidney development.